ZIC5在前列腺癌中的表达及意义

Effect of ZIC5 on biological behaviors of prostate cancer and its mechanisms

目的研究ZIC5(Zic family member 5)在前列腺癌中的异常表达及其对前列腺癌生物学表型的影响。方法实时荧光定量聚合酶链式反应(quantitative real-time PCR)检测ZIC5在前列腺癌组织标本和对应癌旁正常组织、PC3和LNCa P细胞株及正常前列腺上皮中的表达水平。siRNA干扰ZIC5后,CCK-8法和Transwell试验检测前列腺癌细胞PC3和LNCa P的增殖、迁移和侵袭。生物信息学分析可能调控ZIC5的miRNA并进行验证。结果 ZIC5在前列腺癌组织中表达水平上升(t=4.2,P<0.001),并且与Gleason评分分级具有相关性(P<0.05),在前列腺癌细胞株中表达水平显著高于正常前列腺癌上皮(P<0.01)。前列腺癌细胞株PC3和LNCa P中siRNA干扰ZIC5 24 h后,细胞增殖减慢,迁移和侵袭能力被抑制。生物信息学及双荧光素酶报告显示,ZIC5受miR-449家族调控,ZIC5与miR-449家族在前列腺癌标本中表达水平呈负相关(ZIC5 vs miR-449a:r=-0.64,P<0.05,ZIC5 vs miR-449b,r=-0.52,P<0.05),双荧光素酶报告基因检测显示miR-449家族结合ZIC5 3’UTR。异常表达的ZIC5可激活EMT通路,干扰ZIC5和过表达miR-449可抑制ZIC5蛋白表达,并逆转前列腺癌细胞中EMT状态。结论 ZIC5受miR-449家族调控,促进前列腺癌生长和侵袭。

Objective To investigate the expression of ZIC5 in prostate cancer and its effect on the biological behavior of prostate cancer. Methods Expression of ZIC5 in prostate cancer samples and prostate cancer PC3 and LNCa P cells were detected by quantitative real-time PCR（ qRT-PCR）. Cell proliferation and migration were determined by CCK-8 and Transwell methods after PC3 and LNCa P cells transfected with ZIC5 siRNA. Bioinformatics was used to predict the upregulated microRNA,and the target gene was verified by dual-luciferase reporter assay. Results Expression of ZIC5 was upregulated in prostate cancer samples compared with adjacent normal tissues（ t = 4. 2,P〈0. 001）,also elevated in prostate cancer PC3 and LNCa P cells（ P〈0. 05）. CCK-8 and Transwell assays showed that cell proliferation,migration and invasion in PC3 and LNCa P cells were inhibited 24 h after transfected with ZIC5 siRNA. Bioinformatics and dual-luciferase assay showed that ZIC5 was the target gene of miR-449 family. The expression level of ZIC5 was negatively correlated with the expression of miR-449 family in human prostate cancer samples（ ZIC5 vs miR-449 a： r =-0. 64,P〈0. 05; ZIC5 vs miR-449 b,r =-0. 52,P〈0. 05）. Over-expression of ZIC5 activated epithelial-mesenchymal transition（ EMT）of prostate cancer cells. Knock-down of ZIC5 and restoration of miR-449 family reversed the ZIC5-activated EMT in prostate cancer. Conclusion Aberrant expression of ZIC5 promotes the growth and invasion of prostate cancer and it is regulated by miR-449 family.